Viral susceptibility,transfection and growth of SPB – a fish neural progenitor cell line from the brain of snubnose pompano,Trachinotus blochii (Lacépède)

This study investigates the susceptibilities of the SPB cell line to fish viruses including giant seaperch iridovirus (GSIV‐K1), red sea bream iridovirus (RSIV‐Ku), grouper nervous necrosis virus (GNNV‐K1), chum salmon reovirus (CSV) and eel herpesvirus (HVA). GSIV‐K1, RSIV‐Ku and CSV replicated well in SPB cells, with a significant cytopathic effect and virus production. However, the cells were HVA and GNNV refractory. To examine the ability of SPB cells to stably express foreign protein, expression vectors encoding GNNV B1 and B2 fused to enhanced green fluorescent protein (EGFP) and GSIV ORF35L fused to DsRed were constructed and introduced by transfection into SPB cells. Stable transfectants displayed different morphologies compared with SPB and with each other. EGFP‐B1 was predominantly localized in the nuclei, EFPF‐B2 was distributed throughout the cytoplasm and nucleus, and granular 35L‐DsRed was localized with secreted vesicles. The expression of EFPF‐B2 in SPB cells produced blebs on the surface, but the cells showing stable expression of EGFP, EGFP‐B1 or 35L‐DsRed showed normal morphologies. Results show the SPB cells and the transfected cells grow well at temperatures between 20 and 35 °C and with serum‐dependent growth. SPB cells are suitable for studies on foreign protein expression and virology.     

Effects of water temperature on mortality in Megalocytivirus‐infected rock bream Oplegnathus fasciatus (Temminck et Schlegel) and development of protective immunity

Rock bream iridovirus (RBIV) causes huge losses, especially in rock bream Oplegnathus fasciatus. Rock bream injected with RBIV and held at 29, 26, 23 or 20 °C had 100% mortality. Conversely, all infected fish held at 17 °C survived even after the temperature was progressively increased to 26 °C at 100 dpi. Rock bream exposed to virus and held for 2, 4 and 7 days at 23/26 °C before the temperature was reduced to 17 °C had mortality rates of 26.6/73.2%, 66.6/100% and 93.4/100%, respectively, through 100 dpi. When surviving fish had the water temperature increased from 17 to 26 °C at 100 dpi, they did not exhibit signs of disease and had low virus copy numbers (below 10³). To investigate the development of a protective immune, rock bream were infected with RBIV and held at 23 °C before shifting the water temperature to 17 °C at 4 dpi. All injected fish survived until 120 dpi. While 100% of the previously unexposed fish died, 80.2% of the previously infected fish survived. When the survivors were rechallenged again at 160 dpi, no further mortality occurred. The high survival rate of fish following rechallenge with RBIV indicates that protective immunity was established in the surviving rock bream.     

Population‐level consequences for wild fish exposed to sublethal concentrations of chemicals – a critical review

Concentrated chemical spills have been shown to impact adversely on fish populations and even cause localized population extinctions. Evaluating population‐level impacts of sublethal exposure concentrations is, however, complex and confounded by other environmental pressures. Applying effect measures derived from laboratory‐based chemical exposures to impacts in wild fish populations is constrained by uncertainty on how biochemical response measures (biomarkers) translate into health outcomes, lack of available data for chronic exposures and the many uncertainties in available fish population models. Furthermore, wild fish show phenotypic plasticity and local adaptations can occur that adds geographic and temporal variance on responses. Such population‐level factors are rarely considered in the chemical risk assessment process and can probably be derived only from studies on wild fish. Molecular technologies, including microsatellite and SNP genotyping, and RNASeq for gene expression studies, are advancing our understanding of mechanisms of eco‐toxicological response, tolerance, adaptation and selection in wild populations. We examine critically the application of such approaches with examples including using microsatellites that has identified roach (Rutilus rutilus) populations living in rivers contaminated with sewage effluents that are self‐sustaining, and studies of stickleback (Gasterosteus aculeatus) and killifish (Fundulus heteroclitus) that have identified genomic regions under selection putatively related to pollution tolerance. Integrating data on biological effects between laboratory‐based studies and wild populations, and building understanding on adaptive responses to sublethal exposure are some of the priority research areas for more effective evaluation of population risks and resilience to contaminant exposure.     

Protein‐sparing capability of dietary lipid in herbivorous and omnivorous freshwater finfish: a comparative case study on grass carp (Ctenopharyngodon idella) and tilapia (Oreochromis niloticus × O. aureus)

Two, 8‐week feeding trials were conducted to compare protein‐sparing capability of dietary lipid in herbivorous grass carp (Ctenopharyngodon idella) and omnivorous tilapia (Oreochomis niloticus × O. aureus). Utilizing a 2 × 3 factorial design, experimental diets containing two levels of crude protein (380 and 250 g kg⁻¹) and three levels of lipid (0, 40 and 100 g kg⁻¹) were formulated for use in both feeding trials. Growth performances showed better response of both fish fed 380 g kg⁻¹ protein diet than those fed 250 g kg⁻¹ protein diet. Despite the dietary protein level, weight gain (WG), specific growth ratio (SGR), feed conversion ratio (FCR) and protein efficiency ratio were much higher (P < 0.05) for grass carp fed 40 g kg⁻¹ lipid diet than those fed 100 g kg⁻¹ lipid diet; however, there were no significant differences in tilapia fed the two diets. The feed intake of grass carp fed lipid‐free diet was the lowest, but it tended to decrease with increase in dietary lipids in tilapia. Lipid retention (LR) was negatively correlated with dietary lipid concentration of both fish. Viscerosomatic index (VSI), hepatosomatic index (HSI), intraperitoneal fat ratio (IPF) and whole‐body and liver lipid content positively correlated with dietary lipid concentration of both fish. Plasma parameters and liver enzymes activities were also positively correlated with dietary lipid concentration of both fish. Liver lipid contents were higher and enzymes activities were lower in grass carp when compared with tilapia. These data suggested that there was no evidence of a protein‐sparing effect of dietary lipids in grass carp. Tilapia has relatively higher capacity to endure high dietary lipid level compared to grass carp.     

Use of chitin and krill in aquaculture – the effect on gut microbiota and the immune system: a review

Chitin consists of β‐1,4‐linked N‐acetylglucosamine residues and is estimated as the second most abundant biomass in the world after cellulose. However, relatively little chitin is utilized as a material for industrial, agricultural and medical applications and aquacultural purposes. Chitin may be useful as a constitutive material in formulated fish feed, and the interesting effects in fish merit further evaluation. There is evidence that fish and aquatic animals harbour a gut bacterial community that is distinctly different from that reported in the surrounding habitat or in the diet. Thus, the gut environment provides a specific niche, and bacterial activity in the gut is not merely a continuum of that observed in the environment. Today, it is well accepted that the gut microbiota in fish are modulated by dietary manipulations. But to what extent can dietary chitin and krill (chitin‐rich) modulate the intestinal microbiota of fish and how do these dietary components affect the immune system? These questions will be discussed in the present review.     

Sawfishes in The Gambia and Senegal – shifting baselines over 40 years

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Piscine reovirus in wild and farmed salmonids in British Columbia,Canada: 1974–2013

Piscine reovirus (PRV) was common among wild and farmed salmonids in British Columbia, western Canada, from 1987 to 2013. Salmonid tissues tested for PRV by real‐time rRT‐PCR included sections from archived paraffin blocks from 1974 to 2008 (n = 363) and fresh‐frozen hearts from 2013 (n = 916). The earliest PRV‐positive sample was from a wild‐source steelhead trout, Oncorhynchus mykiss (Walbaum), from 1977. By histopathology (n = 404), no fish had lesions diagnostic for heart and skeletal muscle inflammation (HSMI). In some groups, lymphohistiocytic endocarditis affected a greater proportion of fish with PRV than fish without PRV, but the range of Ct values among affected fish was within the range of Ct values among unaffected fish. Also, fish with the lowest PRV Ct values (18.4–21.7) lacked endocarditis or any other consistent lesion. From 1987 to 1994, the proportion of PRV positives was not significantly different between farmed Atlantic salmon, Salmo salar L. (44% of 48), and wild‐source salmonids (31% of 45). In 2013, the proportion of PRV positives was not significantly different between wild coho salmon, Oncorhynchus kisutch (Walbaum), sampled from British Columbia (5.0% of 60) or the reference region, Alaska, USA (10% of 58).     

Towards sustainable exhibits – application of an inorganic fertilization method in coral reef fish larviculture in an aquarium

Coral reef fish are collected from the wild and exhibited in aquaria worldwide. Some of the fish spawn in captivity; however, the eggs are usually neglected. In this study, we collected the eggs spawned naturally in the exhibit tanks, hatched and cultured them indoor in 2000‐L fibreglass tanks (initial density = 18 000 egg tank^?1). We applied an inorganic fertilization method commonly used in freshwater fish culture in raising these coral reef fish larvae. We maintained inorganic phosphorus concentration at 100 μg P L^?1 and inorganic nitrogen at 700 μg N L^?1 daily in the fertilized group (n = 4), while the control tanks (n = 4) were fed with rotifers (10 ind mL^?1). Chlorophyll a at particle sizes of both 0.45–20 μm and >20 μm, as well as NH₃‐N, NO₃‐N, and PO₄‐P concentrations were significantly higher in the fertilized group than the control. Zooplankton in the size groups of 10–50 μm (mainly flagellates) and 50–100 μm (mainly ciliates) were abundant (about 10~60 ind mL^?1) during 3–7 days in fertilized tanks. The average larval fish survival rate at 21 day after hatch in fertilized group was consistently higher than the control in two trials. The experiments demonstrated that the inorganic fertilization approach can be successfully adapted for coral reef fish culture in an aquarium to achieve sustainable exhibits.     

Spatial and temporal patterns in summer ichthyoplankton assemblages on the eastern Bering Sea shelf 1996–2007

Larval and early juvenile fishes were sampled from the eastern Bering Sea (EBS) shelf from 2001 to 2005, and in 2007. Data from these collections were used to examine spatial and temporal patterns in species assemblage structure and abundance. The years 2001–2005 were unusual because the EBS water temperature was ‘warm’ compared with the long‐term mean temperature. In contrast, 2007 was a ‘cold’ year. The abundance of the five most numerous taxa at 12 stations common to all years sampled (1996–2005, 2007) were significantly different among years. Larval and early juvenile stage Theragra chalcogramma (walleye pollock), a commercially important gadid, were by far the most abundant fish in all years. Bottom depth alone best explained assemblage structure in most years, but in others, bottom depth and water column temperature combined and percent sea‐ice coverage were most important. Abundance of T. chalcogramma larvae increases with water column temperature until 5°C and then becomes level. Higher abundances of Gadus macrocephalus (Pacific cod) larvae occur in years with the greatest percent sea‐ice cover as indicated by GAM analysis. Larvae of Lepidopsetta polyxystra (northern rock sole) increase in abundance with increasing maximum wind speed, but decrease at a later date during the last winter storm. The data are consistent with the hypothesis that oceanographic conditions, specifically water temperature and sea‐ice coverage, affect the spatial and temporal pattern of larval abundances. In general, ichthyoplankton species assemblages can be important early indicators of environmental change in the Bering Sea and potentially other subarctic seas as well.     

Decadal trends in abundance,size and condition of Antarctic toothfish in McMurdo Sound,Antarctica, 1972–2011

We report the analyses of a dataset spanning 39 years of near‐annual fishing for Dissostichus mawsoni in McMurdo Sound, Antarctica, 1972–2011. Data on total length, condition and catch per unit effort (CPUE) were derived from the > 5500 fish caught, the large majority of which were measured, tagged and released. Contrary to expectation, the length frequency of the McMurdo Sound catch was dominated by fish in the upper two‐thirds of the overall distribution exhibited in the industrial catch for the Ross Sea shelf. Fish length and condition increased from the early 1970s to the early 1990s and then decreased. Fish length positively correlated with Ross Sea ice extent in early spring, a relationship possibly caused by more ice encouraging larger fish to move farther south over the shelf and into the study area. Fish condition positively correlated with the amount of open water in the Ross Sea during the previous summer (Feb), perhaps reflecting greater availability of prey with the higher productivity that more open water brings. Decreasing fish size corresponds to the onset of the fishery, which targets the large individuals. CPUE was constant through 2001 and then decreased dramatically. We hypothesize that this decrease is related to the industrial fishery, which began in the 1996–97 austral summer, and concentrates effort over the ice‐free Ross Sea continental slope. As a result of limited prey choices and close coupling among mesopredators of the region, Antarctic toothfish included, the fishery appears to be dramatically altering the trophic structure of the Ross Sea.     

Surveillance of selected nutrients,additives and undesirables in commercial Norwegian fish feeds in the years 2000–2010

This article presents data on commercial, Norwegian fish feeds from 2000 to 2010, including elements, additives, some selected vitamins and a range of environmental contaminants. Iodine, selenium and vitamin D concentrations, all declined during the time period; simultaneously, a reduction in marine ingredients in feeds has occurred. Still, the feeds fulfilled known fish nutrient requirements. Arsenic (As) in the feed was fitted with a polynomial regression with peak concentrations in 2004, correlating with fishery landings of blue whiting (Micromesistus poutassou), a reduction species with high content of As. A polynomial regression was also significant for mercury, which peaked in 2005 and was correlated to fish meal (FM) inclusion in the feeds. Residues of the pesticide DDT and its metabolites, chlordane and toxaphene, as well as polychlorinated biphenyls (PCBs), polybrominated diphenyl ethers and sum dioxins and dioxin‐like PCBs, all decreased during this time period. These positive changes in undesirable substances can likely be attributed to the increasing replacement of marine ingredients with plant ingredients, as well as the increased use of South American fish oils rather than North Atlantic ones. On the other hand, cadmium concentrations were twofold higher in South American FMs, and increased in feeds from 2000 to 2010.     

Cross‐neutralization studies with salmonid alphavirus subtype 1–6 strains: results with sera from experimental studies and natural infections

The serological reactivity between strains of each of the six currently genetically defined subtypes of salmonid alphavirus (SAV) was examined by comparison of homologous and heterologous virus neutralization titres on sera from experimentally infected fish. With the exception of the level of SAV subtype 6 neutralization by heterologous sera, good cross‐neutralization was detected between all subtypes, albeit with variation in geometric mean titres when each subtype‐specific serum set was tested against the panel of virus subtypes. A similar pattern was evident with field sera, except that heterologous neutralization of the SAV6 strain was more evident. In only 23% of available pairwise comparisons was the homologous titre recorded with an experimentally derived serum fourfold or greater than the heterologous titre, and in only two instances was this difference demonstrated in both directions. No virus strains consistently met the old serology‐based criteria (Sub‐committee on Inter‐relationships Among Catalogued Alphaviruses) to be considered separate subtypes within an alphavirus species. Only when testing with an SAV subtype‐2‐specific monoclonal antibody was a major difference between homologous and heterologous neutralization capacity evident. These results provide new direct or indirect information in terms of SAV classification, vaccine efficacy and the selection and validation of reagents for serological and immunological diagnostic purposes.     

Fattv acid content of some freshwater fish in lakes of different trophic levels – a bottom-up effect?

Abstract– Various sizes of roach, perch, pike, zander (pike-perch) and crucian carp were collected from lakes of different trophic levels in order to study the variation of lipids and fatty acids (FA) within and between species. Freeze-dried samples of the dorsal muscle were analysed quantitatively for total lipid content and FA content. The results indicate that total lipid and FA contents can vary considerably, both within and between species. In contrast to herbivorous fish, carnivorous-piscivorous fish FA patterns were more constant and independent of size - fry excepted. Lipid and FA contents of roach from two oligotrophic lakes were significantly higher than in roach from a eutrophic lake. Differences in basic food webs may be responsible for these results. In the oligotrophic lakes, the algal flora was dominated by species classified as high quality food for grazers, e. g. flagellates and diatoms. It was shown in earlier papers that these algal groups contain two long-chained FA of φ 3 type (eicosapentaenoic acid and docosahexaenoic acid) which are used as criteria indicating high nutritional value. Conversely, the eutrophic lake was dominated by blue-greens, a group of autotrophs lacking these long-chained φ 3 FA. Blue-greens have generally been classified as poor food for grazers. (The fatty acids are described by three numbers, x:yφz, where x=number of carbon atoms, y=number of double bonds, and z=position of the first double bond counted from the methyl end of the molecule.)     

Spatial distribution pattern and physical – biological interactions in the larval notothenioid fish assemblages from the Bransfield Strait and adjacent waters

The Bransfield Strait and adjacent waters represent one of the most important areas of larval retention off the Antarctic Peninsula. The species composition of larval fish assemblages has been described in detail in previous surveys carried out in the area, but the role of environmental parameters influencing the spatial distribution of early life stages was poorly known. By applying generalized additive models and multivariate analyses, we evaluated the role of environmental variables in shaping the small‐scale distribution of larval fish and investigated the spatial structure of the larval assemblage. It consisted of a few dominant notothenioid species, such as Champsocephalus gunnari, Lepidonotothen squamifrons, Lepidonotothen larseni, Pleuragramma antarctica and Trematomus scotti, and several other rarely caught species. Sea water temperature, salinity and sampling depth were the most important factors determining the spatial distribution of fish with different relative contributions, together explaining more than 80% of total deviance observed. Species richness was mostly affected by salinity, probably due to the narrow range of salinity preference by the species. Cluster analysis of abundance and presence data identified six and five distinct groups, respectively, each of them with substantial contributions of single or rarely two species. Differences in reproductive strategies of adult populations and spatial distribution of early life stages driven by different larval behaviour in response to environmental factors contribute to maintaining a well‐structured larval fish assemblage, ensuring spatial and food niche partitioning.     

A simple approach for assigning the conservation status of freshwater fish based on their natural variability – a case study from the Iberian Ecoregion

A simple, non‐lethal method for assigning conservation status is proposed based on three conservation classes (favourable, unfavourable/inadequate and unfavourable/bad) to fish species based on their natural population variability. To demonstrate this approach, conservation classes were allocated to three native fish species (brown trout Salmo trutta L., northern Iberian chub Squalius carolitertii (Doadrio), and northern straight‐mouth nase, Pseudochondrostoma duriense (Coelho) based on basic population variables (density and biomass) using a 9‐year dataset (2004–2012) from 15 different sampling sites in Spain. The classification of the population variables into conservation classes was made using previously described nonparametric methods and percentiles categorisation, and the current conservation status of the studied species was established. Temporal trend analyses showed that S. carolitertii experienced a progressive significant decrease during the study period, whereas both S. trutta and P. duriense exhibited weak increases in density. The study exemplifies the need to quantify the natural variability of fish resources to reveal their current conservation status and to determine whether short‐term and long‐term management actions are needed to rehabilitate native fish assemblages. The proposed approach could easily be employed by fisheries managers to monitor the conservational status of other native species.     

Density effects of silver carp Hypophthalmichthys molitrix and catla Catla catla on the production system in all‐male freshwater prawn–finfish polyculture ponds

The effects of three different combinations of silver carp Hypophthalmichthys molitrix and catla Catla catla density on the production system in all‐male freshwater prawn–finfish polyculture ponds were evaluated in triplicate. The stocking density of silver carp and catla, respectively, were maintained at 2000 and 500 ha^?1 in treatment SC₂₀₀₀C₅₀₀, 1500 and 1000 ha^?1 in treatment SC₁₅₀₀C₁₀₀₀ and 1000 and 1500 ha^?1 in treatment SC₁₀₀₀C₁₅₀₀. Male freshwater prawn Macrobrachium rosenbergii and small fish mola Amblypharyngodon mola densities were fixed in all treatments at 12 000 and 20 000 ha^?1 respectively. Management practices were same for all treatments. Blue‐clawed male prawns were harvested twice during the 122‐day culture at 15‐day intervals before the final harvest. Plankton and macro‐benthos abundance and water quality parameters (except transparency and chlorophyll a) did not vary significantly (P>0.05) among treatments. Mean final weights of both silver carp and catla were decreased with the increasing of their own stocking density. The treatment SC₁₅₀₀C₁₀₀₀ resulted in 25–32% increased net production of silver carp plus catla (461 kg ha^?1) and 20–21% increased net production of all species combined (874 kg ha^?1) as compared with the other treatments, although the differences in production of prawn and mola among treatments were not significant.     

Effects of alternative dietary oils on lipid metabolism and related gene expression in hybrid grouper (♀Epinephelus fuscoguttatus × ♂E. lanceolatu)

An 8‐week growth trial was conducted to evaluate effects of dietary oil sources on growth, enzymes activity and genes expression levels related to lipid metabolism of hybrid grouper (♀Epinephelus fuscoguttatus × ♂E. lanceolatu) juveniles. Seven iso‐lipid (97 g/kg of dry matter) and iso‐protein (503.5 g/kg of dry matter) experimental diets were formulated containing 50 g/kg fish oil (FO; acting as controls) or various vegetable oils (VOs): corn oil (CO), sunflower oil (SO), tea oil (TO), olive oil (OO), rice oil (RO) and mixed oil (MO; comprising equal amounts of these oils). Each diet was fed to triplicate groups of 40 fish for per repetition (15.09 ± 0.01 g) for 56 days. The results show that (a) alternative dietary oils had no significant effects on final weight compared with control group (p > .05); (b) compared with FO group, VOs significantly changed the contents of serum lipoproteins, cholesterol, triglycerides and the activity of liver lipid‐metabolizing enzymes (p < .05); (c) CO group had the least effect on the serum lipoproteins, triglycerides and cholesterol of grouper compared with control; the activity of liver lipid‐metabolizing enzymes in RO and control group was the closest; (d) the mRNA levels of Δ6 Fatty acid desaturase (Δ6Fad), hormone‐sensitive lipase (HSL) and lipoprotein lipase (LPL) were not significantly effected by lipid sources, but CO, TO, OO and MO significantly down‐regulated the expression of fatty acid synthetase (FAS) mRNA level in liver, while RO opposite (p < .05); (e) vegetable oil significantly up‐regulated peroxisome proliferator‐activated receptor α (PPARα) and peroxisome proliferator‐activated receptor β (PPARβ) mRNA levels, while TO and RO down‐regulated peroxisome proliferator‐activated receptor γ (PPARγ) mRNA levels (p < .05); and 6) MO significantly increased the mRNA levels of heart‐type fatty acid‐binding protein (H‐FABP) and adipocyte‐type fatty acid‐binding protein (A‐FABP) (p < .05), while other VOs had no effect on them (p > .05). In conclusion, dietary substitution of FO by VO in diet affected lipid metabolism of grouper, which may be regulated by PPARs.     

Otolith geochemistry in young‐of‐the‐year peacock bass Cichla temensis for investigating natal dispersal in the Rio Negro (Amazon – Brazil) river system

This study examined otolith geochemistry as a natural marker of natal origins in young‐of‐the‐year (YOY) Cichla temensis in the Negro River Basin of Brazil. We analysed trace element and isotopic composition of otoliths of YOY collected off spawning nests from the main stem and major tributaries. These were compared with regional bedrock geologic composition to explore underlying mechanisms of differences in otolith geochemistry. Our results suggest that spatial differences in otolith geochemistry can be used to distinguish natal origins based on ⁸⁷Sr/⁸⁶Sr, Sr/Ca and Ba/Ca ratios. This approach allowed us to correctly classify 99% of juvenile fish to their natal streams using cross‐validation in a linear discriminant function analysis (LDFA). Patterns of otolith isotopic composition correspond with patterns in regional geology as expected based on previously demonstrated correlations, although some fine‐scale spatial differences cannot be accounted for by available geologic information. These results demonstrate that otolith chemistry is valuable as a natural marker of natal origins in this system and suggest that inferences from geologic maps may be useful for interpreting movements based on otolith geochemical signatures. This information provides the basis for future work to investigate the early life history and spatial ecology of this important cichlid.     

Effect of treatment of chum salmon Oncorhynchus keta (Walbaum) eggs with 1,3;1,6‐β‐D‐glucans on their development and susceptibility to Saprolegnia infection

The effects of six 1,3;1,6‐β‐D‐glucooligo‐ and polysaccharides with different structures (ranging from 1 to 10 kDa in molecular mass and containing 10–25% of β‐1,6‐linked glucose residues) from brown algae, Saccharina cichorioides, on development of the chum salmon, Oncorhynchus keta (Walbaum), were evaluated. Exposure of chum salmon eggs to 1,3;1,6‐β‐D‐glucans with a molecular mass of more than 2 kDa increased the survival of embryos and juveniles and their resistance to Saprolegnia infection by up to 2.5‐fold, leading to a weight gain in juveniles of 40–55% compared with The control chum salmons. The 1,3;1,6‐β‐D‐glucans with molecular mass of 6–8 kDa and used at a at concentration of 0.5 mg mL^?1 rendered the best stimulative effect.